Cellulase composition for treating cellulose-containing textile material

ABSTRACT

The present invention is directed to a new cellulase composition for treating and finishing cellulose-containing textiles. The improved properties of the cellulase composition are based on an elevated content of EGII endoglucanase type component in an otherwise complete cellulase composition. When the composition with elevated EGII contents is used improved color properties, increased lightness, improved visual appearance and reduced pilling tendencies are achieved. The strength properties of the textile materials are essentially unchanged as compared to previously used cellulase compositions.

BACKGROUND OF THE INVENTION

1. The Technical Field of the Invention

The present invention is directed to a cellulase composition having anelevated content of EGII type cellulase. Said cellulase composition isobtainable from bacteria or fungi, especially from Trichoderma. Theinvention is also related to treatment media containing said cellulosecomposition as well as to methods for treating and finishing textilematerials.

2. The State of the Art

Cellulase treatment of cellulose-containing textile materials duringtheir manufacture or finishing is known per se in the art. The enzymatictreatment for finishing of cellulose-containing textile materials iscalled biofinishing. Biofinishing has been used to remove all kinds ofimpurities and individual loose fibre ends that protrude from thetextile surface. The key benefits offered by biofinishing withcellulases are permanent improval of depilling, cleared surfacestructure by reduced fuzz, improved textile handle, such as softness,smoothness and a silkier feel, improved drapability and brighter colorsof the textile and an improved capability of absorbing moisture.

Additionally, cellulases have been used to impart a stone-washedappearance to denims. Complete biodegradation of cellulases is anadvantage of cellulase treatment, which consequently stands out as anenvironmentally-friendly alternative for chemical treatment.

Strength loss of the fabric is a problem associated with cellulasetreatment. The strength loss is caused by cellulase-induced hydrolysisof beta-1,4-glucosidic bonds of cellulose, which in turn results inpartial degradation of cellulosic polymer, and further, can result instrength loss of the fabric.

In the treatment of fabrics, cellulase derived from fungi, for examplefrom Trichoderma reesei, is generally employed, such cellulase beingcomposed of cellobiohydrolase (CBH), endoglucanase (EG) andbeta-glucosidase (BG) type components. The CBH and EG components canfurther be divided into CEHI and CBHII types and into several various EGtypes, the main types of the latter being EGI and EGII. The BGcomponents do not react with cellulosic polymers, but further cleave thedegradation products, for example cellobiose, that are formed as aresult of the synergistic effect of the CBH and EG components.

The isolation of cellulase components from fungi is known in the art.See, for example, Bhikhabhai, R. et al., (1984), Saloheimo, M. , et al.,(1988) Wood et al., (1988), Bhat, K. M. et al., (1989) and Schulein(1988).

A method for treating cotton fabrics, prior to dyeing and finishing,with cellulose solution in order to remove lint and loose surface fibresto impart a better appearance to the fabric is disclosed in the U.S.Pat. No. 5,232,851. The employed cellulase can be produced, for example,by Trichoderma reesei, T. koningii, Penicillium sp or Humicola insolensspecies. In the given examples, CYTOLASE 123 -cellulase (Genencor Int.)was used with no detailed composition given in said publication. Inaddition to cellulose, the cellulose solution may contain buffers,surfactants, abrasion agents, and the like. After treatment, the tensilestrength of the cotton woven fabric was reported to be at least 50percent of the tensile strength of untreated fabric.

The original cellulase composition obtainable from the fermentationmedium, which is derived directly from the microbes, for example from T.reesei fungi, is seldom suitable as such to give a desired result. Theoriginal cellulase composition may comprise about 45-80% CBHI, 10-25%CBHII, 5-15% EGI and 8-15% EGII of the total cellulase protein content.The interrelations of the CBH and EG components contained in acomposition may hence be changed by various methods known to personsskilled in the art. Such methods include, for example, fractionation andgenetic engineering.

The U.S. Pat. No. 5,120,463 for example discloses a detergentcomposition comprising a surfactant and additionally 0.002 to 10 weightpercent of cellulase composed of CBHI type and EG type components withthe ratio of CBHI to (EGI+EGII) being ≧10:1.

The International Patent Application WO 93/22428 discloses a method fortreating cotton fabrics with fungal cellulase compositions comprisingCBHI type and EG type components in a weight ratio greater than 10:1.The application mentions that the tensile strength of treated fabric isat least about 50 percent of the tensile strength of untreated fabric.No test results, however, are given.

In the International Patent Publication WO 94/23113 a method fortreating cotton-containing and non-cotton cellulosic fabrics duringmanufacture to reduce lint generation is described. The cellulasecompositions contain all EG type components to all CBH type componentsin a ratio of greater than 5:1.

In the International Patent Publication WO 92/06221 cotton-containingfabrics treated with a cellulase solution essentially free from CBHItype cellulase components are said to have a decreased strength loss ascompared to fabrics treated with cellulase solution containing acomplete cellulase composition.

Cellulases have been employed also in the finishing of denim fabrics ordenim garments, in order to impart a stone-washed appearance to thefabric.

The stone-wash was traditionally performed using so-called pumicestones. However, the use of pumice stones causes laundries severalproblems, such as the heaviness of handling the stones, the laboriouspicking by hand of the stones from among the garments, significant wearto the machines with resulting high repair and investment costs, thegrowing amounts of waste caused by broken stones and, additionally, thecomplicated access to pumice stones, as the mining of pumice stones isforbidden in certain countries on environmental grounds.

A method for imparting a stone-washed appearance to denim garments bycellulase enzymes is disclosed, for example, in the U.S. Pat. No.4,832,864. Also in this case, the problem is the weakening, i.e.strength loss, of denim as a result of sole enzyme treatment employed toimpart a stone-washed appearance to the fabric.

It is also known in the art that the activity of the cellulasecomposition depends on the acidity of the environment in which thetreatment is carried out. Most generally, the activity is at its highestat slightly acid pHs, even though compositions functioning in a neutralenvironment may also be employed, and even such cellulase compositionsare known to act in alkaline conditions. However, compositions used in aneutral or alkaline environment have a prolonged reaction time, i.e. thecompositions are acting slower. Because time means money, time consumingtreatments are not only less time-effective but also lesscost-effective. Alternatively, more equipment is required to treat thesame amount of fabrics in the same time. This is also expensive andrequires additional space and facilities. Naturally, buffers known topersons skilled in the art are used to adjust the acidity of cellulasetreatment media, but it does not solve all the problems.

The problem of obtaining improved color retention/restoration propertiesas well as improved softening and feel and visual appearance propertiesto cotton fabrics has been discussed in several patents and patentapplications.

In the U.S. Pat. No. 5,090,474, for example, a detergent compositioncontaining substantially pure EGIII is described. The compositioncontains no more than 5 weight percent of CBHI type components.

The International Patent Publication WO 92/06210 describes a cellulasecomposition, which is enriched with unspecified endoglucanase typecomponents. The ratio of EG type components to all CBHI type componentsis greater than 5:1. The composition is said to impart improvedsoftening properties as well as retention/restoration properties to thedetergent composition when used in acidic, neutral or alkaline washingmedia.

U.S. Pat. No. 5,246,853 discloses an improved method for treatingcotton-containing fabrics with fungal cellulase compositions which areessentially free from CBH I type cellulase components, but contains atleast 10% of unspecified EG type components based on the total weight ofproteins in the cellulase solution.

In the International Patent Publication WO 92/06165 a cellulasecomposition is disclosed which composition contains 0.01-5 weightpercent cellulase components. The compositions contains one or moreunspecified EG type components and of all cellulase components less than5 weight percent is of CBH I type components.

The International Patent Publication WO 95/25840 describes a cellulasecomposition, which is essentially free from CBH I type cellulasecomponents and has a weight ratio of all EG-type components to all CBH Itype components of greater than 5:1. Said International PatentApplication corresponds to U.S. Pat. No. 5,525,507, which discloses acomposition useful for treating non-cotton material. The composition isessentially free of all CBHI type cellulase components and the improvedproperties are achieved by modifying the naturally complete fungalcellulase composition by adding at least 10 percent of unspecifiedendoglucanase components.

The International Patent Publication WO 92/17572 describes a cellulasecomposition, which is essentially free from CDH I type cellulasecomponents and comprises at least about 20 weight percent of unspecifiedEG type components.

The International Patent Publication WO 92/17574 describes a cellulasecomposition, which comprises one or more unspecified EG component andone more CBHI component and the ratio of all EG to all CBHI componentsis greater than 5:1.

in view of the above, the primary problem of using cellulasecompositions in the treatment and finishing of cellulose-containingtextile materials has not only been the strength loss of the fabric as aresult of the cellulose treatment but also the fact that even if theproportions of CBH and EG type components have been changed in the mostsubtle ways as suggested in the prior art patents and patentapplications no significant change in the biofinishing properties hasbeen achieved. It seems as if the different ratios of CBH and EG typecomponents are varied more or less randomly and the results, such asremoval of impurities and individual loose fibre ends that protrude fromthe textile surface are as randomly obtainable. The key benefits claimedby the patents and patent application using different ratios of EG andCBH type components in biofinishing, i.e. permanent improval ofdepilling, cleared surface structure by reduced fuzz, improved textilehandle, such as softness, smoothness and a silkier feel, improveddrapability and brighter colors of the textile and improved moistureabsorbability seems to be essentially unchanged even if the ratios of EGand CBH vary. Furthermore, the removal of certain components andaddition of other components to the natural complete fungal cellulasecomposition is not necessarily sufficiently cost-effective.

We have now surprisingly found that it is not the different ratios ofCBH components and EG components that gives the improved properties inmanufacturing and finishing of textiles. It is an increased level ofEGII which is responsible for the improvement. The same or even betterresults in manufacturing and finishing are obtained as soon as evensmall amounts of EGII type component(s) are added to the naturalcomplete cellulase composition background. This improvement is totallyindependent of the ratios of CBH and EG disclosed in prior art.

Thus, the objective of the present invention is to provide an improvedcellulase composition for treating cellulose-containing textilematerials that gives a smooth feel, improved appearance and softness aswell as permanent depilling to the textile.

In particular, the objective of the present invention is to provide animproved cellulase composition for treating cellulose-containing textilematerials that would not result in significant strength loss of thetextile as a result of the treatment.

Additionally, the objective of the present invention is to provide animproved cellulase composition for imparting an acceptable stone-washedappearance to denims and denim garments without causing significantstrength loss to the denims or denim garments.

Further, the objective of the present invention is to provide animproved treatment medium for cellulose-containing textile materialsthat may comprise, in addition to the cellulase composition of thisinvention, for example, surfactants, polymers, buffers, bulk agents,preservatives, stabilizers and/or abrasion agents.

The objective of the present invention is to provide compositions with adecreased reaction time, i.e. compositions which act more rapidly. Thismeans more time- and cost-effective treatment procedures and savings inequipment as well as treatment facilities.

The objective of the present invention is additionally to provide animproved method for treating cellulose-containing textile materials soas to preserve the strength properties, as well as to obtain an improvedappearance and smooth feel of the textile.

A most import objective of the present invention is to carry out theproduction more cost-effectively. The composition of the presentinvention is cheaper, because less activity is required, because theenzyme is more effective than other enzymes. Thus, the amount requiredin the treatment medium is also smaller.

A further objective is to obtain a color tones which are pleasant to thecustomers. It has been shown that greyish tones which are especiallypleasant to the eye and appreciated by customers are obtainable with thecomposition of the present invention.

The described objectives of the present invention have been achieved byemploying a cellulase composition which comprises elevated amounts ofEGII type cellulase as compared to the complete cellulase compositionsas well as other cellulase compositions known from prior art.

The improved cellulase composition of the present invention isobtainable from a cellulase solution produced by cultivating fungi orbacteria and collecting the spent essentially cell-free fermentationbroth or medium containing the naturally occurring complete backgroundcellulase components such as CBH and EG components, but above all anelevated content of EGII components as compared to the natural completecellulase compositions and the prior art cellulase compositions withdifferent CBH and EG ratios. The elevated content of EGII should be suchthat the desired effects are achieved. The EGII type cellulase being thecomponent which is essential for obtaining the objectives and benefitsof the present invention.

THE SUMMARY OF THE INVENTION

The present invention is related to an improved cellulase compositionfor treating cellulose-containing textile materials during theirmanufacture or for their finishing. The improved cellulase compositioncomprises a desired, at least somewhat elevated content of EGII typecomponents as compared to the natural complete cellulase compositionobtainable from a cell-free fermentation medium. When such cellulasecompositions with an elevated content of EGII are used for treatingcellulose containing fabrics the results are improved biofinishingeffects and/or stone-washed appearance and essentially no strength lossin the textile materials as compared with the results obtainable withprior art ratio modified or natural complete cellulase compositions.

The improved cellulase composition of the present invention comprises anelevated content of the EGII type component as compared to previouslyused complete and modified cellulase compositions. The amount of EGIIbeing at least 15-100 weight percent, preferably at least 20-60 weightpercent, most preferably at least 25-40 weight percent of the cellulasecomposition. The ratio of EG:CBH in T. reesei ALKO3529 the EGIIoverproducing strain is estimated to be approximately 0.6-1:1 and theratio of CBHI:EG is estimated to be approximately 1-1.4:1.

The improved cellulase composition of the present invention can also beessentially free from substantially all CBH type components and/or fromsubstantially all EG type component except the EGII type component.

The improved cellulase composition of the present invention isobtainable from fungi or bacteria, especially from fungi such asTrichoderma, Penicillium, Aspergillus, Humicola or Fusarium genera. Themost preferred source of the cellulase composition being a Trichodermareesei species, especially EGII over-producing Trichoderma reeseistrains.

The present invention also provides an improved treatment medium(composition) for treating cellulose-containing textile materials. Saidtreatment medium also contains the improved cellulase composition withthe increased amount of EGII type cellulose as compared to prior artmodified noncomplete and natural complete cellulase compositions. Thecellulase containing treatment medium not only gives improved propertiesduring manufacturing and finishing to the cellulose-containing textilematerials but also has the same or less strength loss when compared withcellulose-containing materials treated under the same conditions withcellulase compositions without the elevated EGII content.

The treatment medium is useful for biofinishing of cellulose-containingtextile materials and for imparting a stone-washed appearance to denimfabrics or garments. Said medium contains surfactants, polymers,buffers, bulking agents, preservatives, stabilizers and/or abrasionagents.

The present invention is also related to a method for treatingcellulose-containing textile materials, wherein the medium defined aboveis used for treating or finishing of cellulose-containing textilematerials. Said modifiable cellulose-containing textile materials arecotton, flax, linen, hemp, ramie, jute, viscose, Polynosic®, Modal,lyocell, Cupro, etc.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows the plasmid map of pALK537.

FIG. 2 shows the plasmid map of pALK540.

FIG. 3 shows the plasmid map of pALK546.

FIG. 4 shows the plasmid map of pALK543.

FIG. 5 shows the lightness of the denim fabrics after 1 hour treatmentin Launder-Ometer with VTT-D-79125 cellulase preparation and withVTT-D-79125 cellulase preparation added with different amounts ofpurified Trichoderma cellulases.

FIG. 6 shows the lightness of the denim fabrics after 2 hours treatmentin Launder-Ometer with VTT-D-79125 cellulase preparation and withVTT-D-79125 cellulase preparation added with different amounts ofpurified Trichoderma cellulases.

FIG. 7 shows the pilling removal from woven cotton fabrics after 1 hourtreatment in Launder-Ometer with different dosages of cellulasepreparations of VTT-D-79125, ALKO3760, ALKO3798, ALKO4097, ALKO2656,ALKO3529 and ALKO3528.

DETAILED DESCRIPTION OF THE INVENTION Definitions

In the description that follows, a number of terms used in textileindustry and enzyme technology as well as recombinant DNA technology areextensively utilized. In order to provide a clear and consistentunderstanding of the specification and claims, including the scope to begiven such terms, the following definitions are provided.

Cellulose-containing textile materials or fabrics

The term "cellulose-containing textile materials or fabrics" as used inthe present invention refers to textile material composed solely orpartly of cellulosic fibres. The textile material includes fibre, yarn,woven fabric, knit, or a ready-made garment, in whose manufacturecotton, flax, linen, hemp, ramie, jute or man-made cellulosic fibres,for example, vis- cose, Polynosic, Modal, lyocell (e.g. Tencel®), Cupro,etc., have been used as raw material. When using synthetic man-madefibres, the amount of cellulosic fibre in the textile material has to beat least 30 percent, preferably over 50 percent.

Complete cellulase composition

The term "complete cellulase composition" as used herein refers tocellulases derived from fungi, for example from Trichoderma reesei. Suchcellulase being composed of cellobiohydrolase (CBH), endoglucanase (EG)and beta-glucosidase (BG) type components. The CBH and EG components canfurther be divided into CBHI and CBHII types and into several various FGtypes, the main types of the latter being EGI and EGII. The BGcomponents do not react with cellulosic polymers, but further cleave thedegradation products, for example cellobiose, that are formed as aresult of the synergistic effect of the CBH and EG components. As onetypical example of such Trichoderma reesei strains the strainVTT-D-79125 is used in the present invention.

The original cellulase composition obtainable from the fermentationmedium, which is derived directly from the microbes, for example from T.reesei fungi having a cellulase composition comprising about 45-80%CBHI, 10-25% CBHII, 5-15% EGI and 8-15% EGII of the total cellulaseprotein content is considered to be the complete cellulase compositionaccording to the present invention. The ratio of EG:CBH in T. reeseiALKO3529 the EGII overproducing strain is estimated to be approximately0.6-1:1 and the ratio of CBHI:EG is estimated to be approximately1-1.4:1.

Modified cellulase composition

The term "modified cellulase composition" as used herein refers to thecomplete cellulase composition in which the interrelations of the CBHand EG components contained in a composition have been changed byvarious methods known to persons skilled in the art. Such methodsinclude, for example, fractionation and genetic engineering as well ascombining different cellulase solutions.

Essentially free from CBH-type components means that at least oneCBH-type component selected from a group consisting of CBHI, CBHII orboth are missing from the cellulase composition, i.e. said compositionis produced by a host from which the genes encoding said proteins areremoved.

Essentially free from EG-type components means that at least one EG-typecomponent selected from a group consisting of EGI, EGIII, EGV or all orany combination of said EG-proteins are missing, i.e. said compositionis produced by a host from which one or more genes encoding saidproteins are removed.

Biostoning

"Biostoning" of fabric or garment means the use of enzymes in place of,or in addition to, the use of pumice stones for the treatment of fabricor garment, especially denim.

Biofinishing

"Biofinishing" refers to the use of enzymes in a controlled hydrolysisof cellulosic fibers in order to modify the fabric or yarn surface in amanner that prevents permanently pilling, improves fabric handle likesoftness and smoothness, clears the surface structure by reducingfuzzing, which results in clarification of colors, improves thedrapability of the fabric, improves moisture absorbability and which mayimprove also the dyeability.

Backstaining

Released dye has a tendency to redeposit on the surface of the fabricfibers. This effect is termed "backstaining."

Detergent

By detergent is meant a cleansing agent that can contain surface activeagents (anionic, non-ionic, cationic and ampholytic surfactants),builders and other optional ingredients such as antiredeposition andsoil suspension agents, optical brighteners, bleaching agents, dyes andpigments and hydrolases. Suitable listing of the contents of detergentsis given in U.S. Pat. No. 5,443,750, a suitable list of surfactants isgiven in U.S. Pat. No. 3,664,961.

Enzyme preparation

By "enzyme preparation" is meant a composition containing enzymes.Preferably, the enzymes have been extracted from (either partially orcompletely purified from) a microbe or the medium used to grow suchmicrobe.

"Extracted from" means that the desired enzymes are separated from thecellular mass. This can be performed by any method that achieves thisgoal, including breaking cells and also simply removing the culturemedium from spent cells. Therefore, the term "enzyme preparation"includes compositions containing medium previously used to culture adesired microbe(s) and any enzymes that have been released from themicrobial cells into such medium during the culture or downstreamprocessing steps.

By a host that is "substantially incapable" of synthesizing one or moreenzymes is meant a host in which the activity of one or more of thelisted enzyme is depressed, deficient, or absent when compared to thewild-type.

By an amino acid sequence that is an "equivalent" of a specific aminoacid sequence is meant an amino acid sequence that is not identical tothe specific amino acid sequence, but rather contains at least someamino acid changes (deletions, substitutions, inversions, insertions,etc) that do not essentially affect the biological activity of theprotein as compared to a similar activity of the specific amino acidsequence, when used for a desired purpose. The biological activity of acellulase, is its catalytic activity, and/or its ability to bind tocellulosic material. Preferably, an "equivalent" amino acid sequencecontains at least 80%-99% identity at the amino acid level to thespecific amino acid sequence, most preferably at least 90' and in anespecially highly preferable embodiment, at least 95% identify, at theamino acid level.

Cloning vehicle

A cloning vehicle is a plasmid or phage DNA or other DNA sequence (suchas a linear DNA) that provides an appropriate nucleic acid carrierenvironment for the transfer of a gene of interest into a host cell. Thecloning vehicles of the invention may be designed to replicateautonomously in prokaryotic and eukaryotic hosts. In fungal hosts suchas Trichoderma, the cloning vehicles generally do not autonomouslyreplicate and instead, merely provide a vehicle for the transport of thegene of interest into the Trichoderma host for subsequent insertion intothe Trichoderma genome. The cloning vehicle may be further characterizedby one or a small number of endonuclease recognition sites at which suchDNA sequences may be cut in a determinable fashion without loss of anessential biological function of the vehicle, and into which DNA may bespliced in order to bring about replication and cloning of such DNA. Thecloning vehicle may further contain a marker suitable for use in theidentification of cells transformed with the cloning vehicle. Markers,for example, are antibiotic resistance. Alternatively, such markers maybe provided on a cloning vehicle which is separate from that supplyingthe gene of interest. The word "vector" is sometimes used for "cloningvehicle."

Expression vehicle

An expression vehicle is a cloning vehicle or vector similar to acloning vehicle but which is capable of expressing a gene of interest,after transformation into a desired host. When a fungal host is used,the gene of interest is preferably provided to a fungal host as part ofa cloning or expression vehicle that integrates into the fungalchromosome, or allows the gene of interest to integrate into the hostchromosome. Sequences that are part of the cloning vehicle or expressionvehicle may also be integrated with the gene of interest during theintegration process. In T. reesei, sites of integration to which thegene of interest can be directed include the cbh and/or the egl loci.Most preferably, the gene of interest is directed to replace one or moregenes encoding undesirable characteristics.

The gene of interest is also preferably placed under the control of(i.e., operably linked to) certain control sequences such as promotersequences provided by the vector (which integrate with the gene ofinterest). Alternatively, the control sequences can be those at theinsertion site.

The expression control sequences of an expression vector will varydepending on whether the vector is designed to express a certain gene ina prokaryotic or in a eukaryotic host (for example, a shuttle vector mayprovide a gene for selection in bacterial hosts). Expression controlsequences can contain transcriptional regulatory elements such as,promoters, enhancer elements, and transcriptional termination sequences,and/or translational regulatory elements, such as, for example,translational initiation and termination sites.

THE GENERAL DESCRIPTION OF THE INVENTION

In the manufacturing or finishing of cellulose-containing textilematerials, it was unexpectedly found that by using a cellulasecomposition, which comprised an increased amount of ECII type cellulasecomponent in an otherwise natural complete cellulase composition, acellulose-containing textile product with essentially the same orimproved properties could be produced. The cotton containing textilematerial also had an improved nice appearance and feel, and showed areduced tendency of pilling, and additionally maintained its strength.Also denims and denim garments treated with the cellulase compositioncontaining elevated amounts of EGII achieved the same or improvedstone-washed appearance as compared to modified and natural completecellulase compositions.

In order to obtain the improved results of the present invention duringmanufacturing and finishing the cellulase composition or treatment mediacontaining compositions, should have more or less elevated EGIIcontents.

It has been shown that using the cellulase composition of the presentinvention improved appearance, softness, drapability, absorption ofmoisture, and brighter colors of cellulose-containing textile materialsis achieved. Also a reduced tendency of pilling and fuzzing is obtained.Additionally, the cellulase composition of this invention can be used inthe finishing of so-called denims to create an improved so-calledstone-washed appearance.

Fungi and bacteria can be used as source material for producing thecellulase composition of the present invention. Preferably the cellulasecompositions of this invention are derived from fungi, for example fromspecies of Trichoderma, Penicillium, Aspergillus, Humicola or Fusariumgenera. Trichoderma reesei being the fungi of preference for producingthe cellulase composition. According to prior art the natural completecellulase compositions comprising CBH components and various EGcomponents, are not as such applicable to the treatment ofcellulose-containing textile materials for producing the desired endresult. The suggested methods of producing compositions which aremodified to give different ratios between the CBH and EG are neithernecessary to achieve the desired results. An elevated EGII-content inthe complete cellulase background composition is the only thing requiredto obtain the desired results.

When some other strain than Trichoderma is used for obtaining thecellulase composition, it is possible, in a similar fashion, to modifythe ratios of the types of cellulase produced by the employed strain,which produces cellulases functionally corresponding to those producedby Trichoderma and adding a suitable amount of EGII, which gives thedesired result.

For a desired and good end result, it is essential that the cellulasecomposition contains a more or less significantly increased amount ofEGII type endoglucanases of Trichoderma. If the cellulase composition isderived from Trichoderma, the cellulase composition of the presentinvention comprises in addition to the elevated EGII-content, EGIcomponents and possibly minor amounts of other EG components, such asEGIII and EGV as well as CBHI and perhaps CBHII components, but saidmore or less undesired EG and CBH components, the ratio of which is ofno major importance, can also be removed using conventional recombinantand fractionation techniques. The ratio of EG:CBH in T. reesei ALKO3529the EGII overproducing strain is estimated to be approximately 0.6-1:1and the ratio of CBHI:EG is estimated to be approximately 1-1.4:1.

The increased EGII content can be provided in cellulase compositionswith a relative ratio of CBHI or EGI to other components increased ordecreased by methods known per se in the art, but as discussed above itis no prerequisite for obtaining the desired results. In the cellulasecomposition of the present invention the ratios of the weights of CBHand EG components are not crucial. It is more important that the amountof EGII type components in the composition is more or less significantlyelevated over the normal level in a complete cellulase compositions.This means that the proportion of EGII should be at least 15-100 weightpercent of the total weight of the complete composition of cellulaseproteins. More preferably, the amount of EGII should be at least 20-60weight percent, and most preferably at least 25-40 weight percent of thecellulase composition. Naturally, the cellulase composition can comprisealmost 100 weight percent of EGII of the total cellulase content, butthis is in no way the preferred embodiment and not necessarily requiredfor obtaining the desired results. The preferred mode of the presentinvention is a natural complete background cellulase composition withelevated EGII-content.

Enzyme activity is one concept applied in determining the properties ofenzymes. The enzyme activities used in the present patent applicationare ECU, FPU and MUL, which are defined as follows:

ECU

The endo-1,4-beta-glucanase in the sample hydrolyses thehydroxyethylcellulose substrate, and the resulting reducing sugars areassayed spectrophotometrically using a dinitrosalicylic acid reagent(DNS). One unit of endo-1,4-beta-glucanase is defined as the amount ofenzyme-producing one nmole of reducing sugars as glucose in one second(1 ECU=1 nkat), (Bailey, M. and Nevalainen, H., 1981).

FPU

The cellulase in the sample hydrolyses the filter paper used as asubstrate, and the resulting reducing sugars are assayedspectrophotometrically using a DNS reagent. The filter paper degradingactivity is described as FPU units. The calculation is based on thedefinition of the International Unit (IU). 1 IU=1 micromol min.-¹ ofproduct formed (reducing sugars as glucose) (IUPAC, 1984).

MUL

The cellobiohydrolase (CBHI) and endoglucanase (EGI) of the samplehydrolyse the 4-methylumbelliferyl-beta-D-lactoside that acts as asubstrate, whereby methylumbelliferone is released that can be measuredspectrophotometrically. The method can be applied in the determinationof cellobiohydrolase I (CBHI) activity. The method also measures theendoglucanase I (EGI) activity, the proportion of which can bedetermined by inhibiting the activity of cellobiohydrolase using 5 mM ofcellobiose. One MUL unit is the amount of enzyme activity that in onesecond under the determination conditions, releases 1 nmol ofmethylumbelliferone from 4-methylumbelliferyl-beta-D-lactoside (vanTilbeurgh et al., 1988).

Treatment of cellulose-containing textile materials with the cellulasecomposition of the present invention imparts a smooth feel, depilling,softness and good appearance to the textiles. The treatedcotton-containing fibres treated with the composition of this inventionhave kept their structure as well as or better than cellulasecompositions comprising prior or conventional cellulase compositions.

It has also been possible to show that the strength properties ofcellulose-containing textile materials, i.e. bursting strength, breakingstrength and tearing strength have remained as good as in textilematerials treated with the prior art compositions, including thecomplete cellulase composition.

Because the desired effect is obtained with shorter treatment times thestrength properties can be even better than with the prior artcompositions.

Treatment media of cellulose-containing textile materials withcompositions disclosed in the present invention may contain, in additionto enzymes, e.g. surfactants, polymers as for example PV-A and PVPpolymers, buffers as for example citrates, acetates and phosphates forregulating the acidity of the solutions, possibly bulk agents,conventional preserving agents, stabilizers and abrasion agents.

The dosage of cellulase products in a solution depends on the desiredresult, application, the activity of the cellulase product etc. Suitablecellulase dosages as disclosed in the present invention, correspond tothe dosages of typical commercial liquid cellulases, as for exampleEcostone L, Ecostone L 20, Biotouch L (Primalco Ltd, Biotec, Nurmijarvi,Finland). The suitable dosages thus fall within the range of 0.05 to 15percent, more preferably within 0.5 to 6 percent of the weight of thetextile material being treated.

The suitable enzyme dosages for imparting biofinishing treatment totextile materials depend on the desired result, on the treatment methodand the activity of the enzyme product. The dosages are about 0.05 to 10percent, more preferably about 0.5 to 5 percent of the weight of thetreated textile material, these dosages corresponding to the dosages oftypical commercial liquid cellulases, as for example Biotouch L,Biotouch C 601, Ecostone L 20 or Ecostone C 80 (Primalco Ltd, Biotec,Nurmijarvi, Finland).

For imparting a stone-washed appearance to denims, the cellulasecompositions of this invention can be successfully employed to replacethe use of pumice stones. The result is a high-quality stone-washedappearance and essentially the same or better strength properties thanthose achieved in fabrics treated with textile treatment mediacontaining no elevated amounts of EGII components.

The suitable enzyme dosages for imparting a stone-washed appearance tothe fabric depend on the desired result, on the treatment method, and onthe activity of the enzyme product, and are about 0.05 to 5 percent,more preferably about 0.5 to 2 percent of the weight of the treatedfabric, these dosages corresponding to the dosages of typical commercialliquid cellulases, as for example Ecostone L, Ecostone L 20, Ecostone LPlus (Primalco Ltd, Biotec, Nurmijarvi, Finland).

The pH range for applying the cellulase composition of this invention isdependent on the pH activity profile of the enzyme. When enzymesfunctioning at acid pHs are being employed, the pH of the applicationenvironment is preferably within the range of 3.5 to 7, more preferablywithin the range of 4 to 5.5.

The cellulase composition of the present invention, which containsconsiderable amounts of Trichoderma EGII type endoglucanases, can beproduced by e.g. fractionation or mutation of the used productionstrain, or by genetic engineering. A cellulase composition with elevatedamounts of EGII type endoglucanases can be prepared correspondingly. Themost convenient way of producing a product with increased amount of EGIIprotein is to add copies of the gene encoding the EGII protein to theproduction strain, e.g. to the Trichoderma strain, as described inExample 1. The selected Trichoderma strain or other employed productionorganism can be a wild type strain, or a strain more suitable for use asa production organism developed from the wild type strain by furthermutation or genetic engineering. As for Trichoderma reesei, suitablestrains include, for example, the wild type strain T. reesei QM6a andthe derived mutant strains, e.g. QM9414 and RutC-30, developed forcellulase production, and strains further developed from these, in whiche.g. the cellulase and/or hemicellulase level has been further raised,and/or in which the level of produced protease has been lowered.VTT-D-79125 and ALKO2221 (a mutant strain with a low level of proteaseproduction) and their derivatives, as for example the strainoverproducing the EGII enzyme, whose construction is disclosed inExample 1, are examples of strains that have been further developed. Insaid strains one or more gene coding for CBH or EGs others than EGII canbe replaced with a gene encoding EGII.

A general description of the methods for inserting the gene encoding theEGII protein, and of the method for increasing the production level ofthe EGII protein in Trichoderma reesei is described in the following.Applying the same method, it is possible to change the ratios ofcellulases also in other species and, for example, to add copies ofgenes encoding Trichoderma EGII type cellulase(s) or to replace somenative genes with genes encoding Trichoderma EGII type cellulase(s).

The gene encoding the EGII protein (egl2; Saloheimo et al., 1988; in thepublication the gene egl2 is referred to as egl3, which is the originalname of the gene) can be inserted into the selected Trichoderma strainor for example, replace a gene encoding EGI (egl1; Penttila et al.,1986), or another gene encoding a not required protein or by simplyadding the copy number by insertions without deleting anything. When theegl2 gene replaces, e.g. the egl1 gene, the EGII activity of the enzymemixture produced by the strain is higher than in the enzyme mixtureproduced by strains from which the egl2 gene is deleted or exist only inone copy. Any marker suitable for Trichoderma can be employed as amarker gene, as for example amdS (e.g. from plasmid p3SR2; Kelly andHynes, 1985), hygB (e.g. from plasmid pRLM_(ex) 30; Mach et al., 1994)and ble (e.g. from plasmid pAN8-1, Mattern and Punt, 1988). Forauxotrophic strains, the complementary gene for the auxotrophy concernedcan also be employed as a marker.

The selected marker gene and the egl2 gene, are ligated between the 5'and 3' flanking regions of the target gene forming a targeting plasmid.Using the flanking regions, homologous recombination can occur and thedesired genes can be targeted to give place for an inserted egl2 gene.The principle of gene replacement is described by Suominen et al.(1993). For the targeting frequency to be satisfactorily high, theflanking regions must be long enough, e.g. in Trichoderma at least 1.5kb in length; examples of flanking regions suitable for replacingcellulase genes have been described by Suominen et al. (1993). Thenecessary flanking regions of the cellulase gene can be isolated fromthe Trichoderma gene bank.

Strains of EGII-overproducing phenotypes can be selected from amongtransformants, for example, by analysing the culture media on the basisof their increased endoglucanase activity as compared to the host strain(ECU activity, Bailey and Nevalainen, 1981).

The production level of the EGII protein can be raised in a selectedstrain, for example in the Trichoderma reesei strain (examples ofapplicable T. reesei strains have been given in the foregoing) bymutation or by increasing the copy number of the egl2 gene, as describedin Example 1A. In addition to the cbh1 promoter, also other promoters,suitable for the selected strain, can be used for the expression of theegl2 gene. Any marker suitable for Trichoderma may be used as a markergene in the transformation, as has been described in the foregoing.

The sources referred to in the above description are given in thereference list in the end of the specification.

The following examples and figures provide further details of thepreparation of the cellulose composition of the present invention, andthe effect of the cellulase composition on the properties ofcellulose-containing textile materials.

EXAMPLE 1

Construction of the strains

Trichoderma reesei strains overproducing each of the main Trichodermacellulase, endoglucanases I and II (EGI and EGII) and cellobiohydrolasesI and II (CBHI and II), were constructed for the production of differentcellulase preparations.

A. Construction of EGII overproducing Trichoderma reesei strain

In construction of the Trichoderma reesei EGII overproducing strain, theparental T. reesei strain VTT-D-79125 was transformed with theexpression cassette from the plasmid pALK537 (FIG. 1). In the cassetteEGII is expressed from the strong cbh1 promoter. T. reesei VTT-D-79125is a hypercellulolytic mutant strain that contains all the mainTrichoderma cellulases (Nevalainen, 1985).

The plasmid pALK537 contains:

* T. reesei cbh1 (cellobiohydrolase 1) promoter from Trichoderma reeseiVTT-D-80133 (Teeri et al., 1983). The 2.2 kb EcoRI-SacII fragment wasused in the construct. The sequence of the promoter area preceeding theATG was published by Shoemaker et al. (1983). The last 15 nucleotides ofthe T. reesei L27 cbh1 promoter (the SacII site is underlined) areCCGCGGACTGGCATC (SEQ ID NO: 1) (Shoemaker et al., 1983). The cbh1promoter from T. reesei strain VTT-D 80133 has been sequenced in thelaboratory of Primalco Ltd. and one nucleotide difference in the DNAsequence has been noticed within the above mentioned region. In the T.reesei VTT-D-80133 the sequence preceeding the ATG is CCGCGGACTGCGCATC(SEQ ID NO:2).

The nucleotides missing from the promoter (10 bps after the SacII to theATG) were added and the exact promoter fusion to the signal sequence ofeg12 (endoglucanase 2) cDNA was done by using the PCR (polymerase chainreaction) method. The fusion and the PCR fragment were sequenced toensure that no errors had occurred in the reaction.

* T. reesei eg12 (endoglucanase 2, originally called eg13) cDNA. Thenucleotide sequence of the eg12 cDNA is described in Saloheimo et al.(1988). In the plasmid pALK537 a 1.4 kb fragment (from the signalsequence to EcoRI site) was used.

* T. reesei cbh1 terminator from T. reesei VTT-D-80133 (Teeri et al.,1983). The 739 bp AvaII fragment starting 113 bp before th STOP codon ofthe cbh1 gene was added after the eg12 cDNA.

* cbh1 3'-fragment: The 1.4 kb BamHI-EcoRI fragment was isolated from T.reesei ALKO2466 (Suominen et al., 1993). 3'-fragment can be usedtogether with the promoter area to target the eg12 cDNA to the cbh1locus by homologous recombination.

* amdS gene: The gene has been isolated from Aspergillus nidulans and itis coding for acetamidase (Hynes et al., 1983) Acetamidase enables thestrain to grow by using acetamide as the only nitrogen source and thischaracteristics has been used from selecting the transformants. The 3.1kb SpeI-XbaI fragment from the plasmid p3SR2 (Kelly and Hynes, 1985) wasused in the pALK537 plasmid.

The standard DNA methods described by Maniatis et al. (1982) were usedin construction of the vectors. The restriction enzymes, T4 DNA ligase,Klenow fragment of DNA polymerase I and alkaline phosphatase from calfintestine used in the DNA manipulations were from Boehringer (Germany)and New England Biolabs (USA). Each enzyme was used according to thesupplier's instructions. Plasmid DNA from E. coli was isolated by usingthe Qiagen colums (Diagen GmbH, Germany) according to the supplier'sinstructions. DNA fragments for cloning or transformations were isolatedfrom low melting point agarose gels (FMC Bioproducts, USA) by the freezethaw phenol method (Benson, 1984). The oligonucleotides used in thePCR-reactions and in sequencing reactions were synthetized by a ABI(Applied Biosystems, USA). Sequencing of the fusion was carried out bythe automated sequencer (Applied Biosystems 373A, USA) T. reeseiVTT-D-79125 was transformed with the 9.2 kb linear NotI fragment ofpALK537 as described by Penttila et al. (1987). T. reesei transformantswere transferred on a selective medium and purified through conidia. Thepurified transformants were grown in shake flasks in a medium containing4% whey, 1.5% complex nitrogen source derived from grain, 1.5% KH₂ PO₄and 0.5% (NH₄)₂ SO₄, pH 5.5. Cultures were grown at 30° C. and 250 rpmfor 7 days. The activity against hydroxyethylcellulose (HEC) wasmeasured from the culture media of the transformants. In thetransformant ALKO3529 the activity against HEC (3400 ECU/ml) was about 3fold compared to the parent strain VTT-D-79125 (1200 ECU/ml). Accordingto Southern blot analysis ALKO3529 strain has at least two tandem copiesof the transformed vector fragment containing the eg12 expressioncassette.

ECU: One unit of ECU activity (endo-1.4-beta-glucanase) is defined asthe amount of enzyme producing one nmole of reducing sugars as glucosein one second from the substrate hydroxyethylcellulose (HEC) 1 ECU=1nkat (Bailey and Nevalainen, 1981).

B. Construction of EGI overproducing T. reesei strain

Construction of T. reesei EGI overproducing ALKO2656 strain, where theCBHI gene has been deleted, is described in Karhunen et al. (1993).

C. Construction of T. reesei strain that overproduces EGI and EGIIwithout CBHI and CBHII

For overproduction of T. reesei EGI and EGII without CBHI and CBHII, theexpression cassette from the plasmid pALK540 (FIG. 2) was transformed toT. reesei ALKO2698 (Karhunen et al., 1993). ALKO2698 is an EGIoverproducer that does not contain the cbh1 gene. In the cassette ofpALK540 EGII is expressed from the strong cbh1 promoter.

The plasmid pALK540 contains:

* T. reesei cbh1 promoter, terminator and eg12 cDNA as described inexample 1A.

* ble gene. The gene has been isolated from Streptoalloteichushindustanus (Drocourt et al. 1990). The gene confers resistance tobleomycin and the related antibiotics, phleomycin and tallysomycin.Resistance to phleomycin has been used for selecting the transformants.3.3 kb XbaI-Bg12 fragment from the plasmid pAN8-1 (Mattern et al., 1987)was used in pALK540. The fragment contains phleomycin gene (ble),flanked by the Aspergillus nidulans gpdA promoter and trpC terminatorregions.

* T. reesei cbh2 5'-fragment. The fragment was isolated from T. reeseistrain VTT-D-80133 (Teeri et al., 1987). A 3.4 kb XhoI-PvuII fragmentstarting 1.4 kb upstream from the cbh2 gene was used to target togetherwith cbh2 3'-fragment the expression cassette to the cbh2 locus.

* T. reesei 3'-fragment. The fragment was isolated from T. reesei strainVTT-D-80133 (Teeri et al., 1987). A 1.6 kb XbaI-BglII fragment starting1.1 kb downstream from the cbh2 gene was used to target together withcbh2 5'-fragment the expression cassette to the cbh2 locus.

The same methods as described in example 1A were used in construction ofthe vectors.

T. reesei ALKO2698 was transformed with the 11.6 kb ClaI-PvuI fragmentof pALK540 as described by Penttila et al. (1987). The transformedprotoplasts were plated onto surface of MnR plates osmoticallystabilized with 0.44M saccharose and incubated for 6 h at 30° C., priorto the addition of 5 ml of molten MnR as an overlay containing 300 μg/mlphleomycin (Cayala, France). The transformants were purified onselective MnR-medium supplemented with 50 μg/ml of phleomycin throughsingle spore before transferring to MnR-slants containing 50 μg/mlphleomycin for three generations and after that to PD slants. Thepurified transformants were grown on microtiter plates for detection ofthe CBHII protein by Western blotting with CBHII monoclonal antibody.ALKO3528 was one of the CBHII-negative transformants and it contains onecopy of the eg12 expression cassette in the place of cbh2 (Southernblot). ALKO3528 transformant was cultivated as in example 1A formeasuring the activity against HEC (ECU/ml) from the culture medium. Theactivity against HEC was increased about 2 fold in ALKO3528 straincompared to the parent strain ALKO2698 and by 4 fold compared to theVTT-D-7912S strain which is a parent strain of ALKO2698.

D. Construction of CBHI overproducing T. reesei strain

Construction of T. reesei CBHI overproducing ALKO3760 strain isdescribed in the International Patent Application Publication WO96/34945 herewith incorporated by reference.

E. Construction of CBHII overproducing T. reesei strain

For overproduction of T. reesei CBHII expression cassette from Theplasmid pALK546 (FIG. 3 ) was transformed to T. reesei ALKO2221(VTT-D-79125 mutant strain possessing a low level of protease activity).In the cassette of pALK546 CBHII is expressed from the strong cbh1promoter.

The plasmid pALK546 contains:

* T. reesei cbh1 promoter and terminator as described in example 1A.

* cbh2 gene. The cbh2 gene was from T. reesei VTT-D-80133 (Teeri et al.,1987). The fragment containing the cbh2 gene is about 2.2 kb in length,1.5 kb of which is coding area.

* ble gene, gpdA promoter and trpc terminator as described in example1C.

* eg12 5'-fragment was isolated from T. reesei strain VTT-D-79125 and itis subcloned from a λ clone, originally called eg13, isolated bySaloheimo et al. (1988). pALK546 contains the 1.4 kb XhoI-SacI fragmentabout 2.2 kb upstream from the eg12 gene. This fragment can be usedtogether with the eg12 3'-fragment to target the expression cassette tothe eg12 locus.

* eg12 3'-fragment. The 1.6 kb AvrII-SmaI fragment about 0.2 kb from theend of the eg12 gene was used in pALK546. The fragment originates fromthe same clone as the 5'-fragment.

pALK546 was constructed according to examples 1A and 1C.

T. reesei ALKO2221 was transformed with the 11.8 kb EcoRI-BamHI fragmentof pALK546 accordingly to example 1C. The purified transformants werecultivated as in example 1A and the amount of secreted CBHII cellulasewas quantitated by ELISA method (Buehler, 1991) with CBHII monoclonalantibody. In the transformant ALKO3798 the amount of CBHII protein wasincreased 4 fold compared to the parental strain ALKO2221. According toSouthern blot analysis ALKO3798 contains one copy of the cbh2 expressioncassette.

F. Construction of T. reesei strain that overproduces CBHI and CBHIIwithout EGI and EGII

For overproduction of T. reesei CBHI and CBHII without EGI and EGIIexpression cassette from the plasmid pALK543 (FIG. 4) was transformed tothe egl2 locus of T. reesei ALKO3761. ALKO3761 is a CBHI overproducingEGI-negative strain. Construction of the ALKO3761 strain is described inWO 96/34945 hereby incorporated by reference, except that theEGI-negative transformants were screened by Western blot with EGImonoclonal antibody. ALKO3761 contains one copy of the expressioncassette in the egl1 locus and the secreted amount of the CBHI cellulaserose compared to the parental strain ALKO2221. In the cassette ofpALK543 cbh2 is expressed under its own promoter.

The plasmid pALK543 contains:

* cbh2 promoter, gene and terminator from T. reesei VTT-D-80133 (Teeriet al., 1987). The 4.7 kb SphI-SacII fragment used in pALK543 containsapproximately 1.5 kb coding area, approximately 2.5 kb promoter regionand approximately 0.7 kb terminator region.

* ble gene, gpda promoter and trpC terminator as described in example1C.

* eg12 5' and 3' fragments as described in example 1E.

pALK543 was constructed according to examples 1A and 1C.

T. reesei ALKO3761 was transformed with the 11.2 kb BamHI fragment ofpALK543 and the transformants were screened accordingly to examples 1Cand 1E. ALKO4097 was one of the EGII-negative transformants andaccording to Southern blot analysis it contains 1 copy of the cbh2expression cassette in the eg12 locus. In ALKO4097 the amount ofsecreted CBHII protein was increased 3 fold compared to the parentalstrain ALKO3761.

EXAMPLE 2

Effects of Trichoderma purified cellulases in biostoning

In this experiment is studied the stone-washing effects of cellulasepreparation derived from Trichoderma reesei VTT-D-79125 strain with theaddition of different purified Trichoderma cellulases CBHI(cellobiohydrolase I), CBHII, EGI (endoglucanase I) and EGII cellulases.VTT-D-79125 is a hypercellulolytic Trichoderma reesei mutant strain,(Nevalainen, 1985).

Denim fabric was prewashed 10 min at 60° C. with Ecostone A 200 (1ml/liter, Primalco Ltd, Biotec, Finland). The fabric was then cut into12×12 cm swatches. The color was measured from the fabric as reflectancevalues with the Minolta (Osaka, Japan) Chroma Meter 1000 R L*a*b*system.

Cellulase Treatments were performed in LP-2 Launder-Ometer (Atlas, Ill.,USA) as follows. About 7 g of denim swatches were loaded into the 1.2liter container that contained 200 ml of 0.05M citrate buffer pH 5. Aquantity of steel balls were added into each container to help the colorremoval. VTT-D-79125 preparation was used as endoglucanase unit (ECU/ml,example 1). 300 ECU per g of fabric was used in each test and purifiedCBHI, CBHII, EGI or EGII cellulases were added 1 or 2 mg per g fabric.The containers were then closed and loaded into a 50° C. Launder-Ometerbath. The Launder-Ometer was run at 42 rpm for 1 and 2 hours.

After removing swatches from the containers they were soaked for 10 minin 200 ml of 0.01M NaOH and rinsed for 2×5 min with cold water. Swatcheswere then dried for 1 hour at 105° C. and air dried overnight at roomtemperature. The color from both sides of the swatches was measured withthe Minolta Chroma Meter. Results from the color measurements of treateddenim fabrics are shown in Table I. FIG. 5 and 6 shows the increase oflightness in the right side of the fabric after 1 and 2 hours treatment.

                  TABLE I    ______________________________________    Color measurements of denim fabrics treated with    VTT-D-79125 cellulase preparation and VTT-D-79125 preparation    added with purified cellulases.    Cellulase    added to    VTT-D-79125         Right side    preparation  mg/g   L          b   deltaE    ______________________________________    1 hour    --*          --     0.8        1.5 1.5    CBHI         1      0.8        2.0 1.5    CBHI         2      1.8        2.0 2.2    CBHII        1      1.1        1.7 1.6    CBHII        2      0.6        1.7 1.3    EGI          1      1.1        2.0 1.9    EGI          2      1.8        2.5 3.0    EGII         1      1.9        3.2 3.2    EGII         2      2.7        2.5 3.8    2 hours    --*          --     1.2        1.9 1.5    CBHI         1      2.2        2.5 3.1    CBHI         2      1.4        2.5 2.7    CBHII        1      1.0        2.2 1.5    CBHII        2      1.3        2.7 2.6    EGI          1      2.2        2.9 2.2    EGI          2      2.0        3.1 3.5    EGII         1      2.5        3.7 1.9    EGII         2      4.2        3.5 5.2    ______________________________________     *pure VTTD-79125 preparation     The effects of the buffer on colors has been deleted from these values.     L: Lightness unit of the fabric after treatment minus lightness unit of     the fabric before the treatment     b: Blueness unit of the fabric after treatment minus blueness unit of the     fabric before the treatment     deltaE: Color difference in the L*a*b* color space between the specimen     color and the target color (target = untreated denim fabric)

The results show that addition of EGII cellulase to the VTT-D-79125preparation increased the lightness (stone-washed effect) of the denimfabrics in stone-washing most compared to the addition of other purifiedcellulases. Addition of CBHI or EGI improved the stone-washing effectcompared to pure VTT-D-79125 preparation but less than with EGII. After1 hour of treatment to obtain the same stone-washed effect a doubleamount of CBHI or EGI compared to EGII was required. Addition of CBHIIgave the same stone-washing effect as pure VTT-D-79125 preparation.

EXAMPLE 3

Effects of different Trichoderma cellulase preparations in biostoning

In this example is studied the stone-washing effects of cellulasepreparations derived from Trichoderma reesei strains ALKO3760, ALKO3798,ALKO 4097, ALKO2656, ALKO3529, ALKO3528 and VTT-D-79125 (example 1).

The experimental set up was as in example 2. 3 and 6 mg of the totalprotein in cellulase preparations per g of fabric was used in eachexperiment. The washing times were 1 and 2 hours at 50° C.

Results from the color measurements are shown in Table II.

                  TABLE II    ______________________________________    Color measurements of denim fabrics treated with    VTT-D-79125, ALKO3760, ALKO3798, ALKO4097, ALKO2656,    ALKO3529 and ALKO3528 cellulase preparations.              dosage                    Right side    preparation mg/g    L         b    deltaE    ______________________________________    1 hour    VTT-D-79125 3       1.5       1.4  1.8                6       1.5       1.8  2.0    ALKO3760    3       1.3       1.2  1.8                6       0.6       1.9  1.6    ALKO3798    3       1.4       1.4  1.4    ALKO4097    3       0         0.8  0                6       0.5       0.8  0.3    ALKO2656    3       0.1       2.6  2.2    ALKO3529    3       1         1.8  1.8                6       2.1       2.9  3.3    ALKO3528    3       1.3       2.0  1.8                6       1.5       2.8  2.3    2 hours    VTT-D-79125 3       1.8       2.3  NM                6       2.4       2.6  NM    ALKO3760    3       2.2       2.9  2.7    ALKO3798    3       1.6       2.5  2.5    ALKO4097    3       0.1       1.2  NM                6       0.8       1.7  NM    ALKO2656    3       2.5       2.9  3.1    ALKO3529    3       3.2       2.6  3.6    ALKO3528    3       2.7       3.2  NM    ______________________________________     NM = not measured     The effects of the buffer on colors has been deleted from these values.     L: Lightness unit of the fabric after treatment minus lightness unit of     the fabric before the treatment     b: Blueness unit of the fabric after treatment minus blueness unit of the     fabric before the treatment     deltaE: Color difference in the L*a*b* color space between the specimen     color and the target color (target = untreated denim fabric)

Results show that after 1 hour treatment with 3 mg/g dosage thestone-washing effect measured as lightness units is almost equal withVTT-D-79125, ALKO3528, ALKO3760, ALKO3529 and ALKO3798 cellulasepreparations. No clear increase in lightness units is obtained withALKO4097 or ALKO2656 preparations. With 6 mg/g dosage after 1 hourtreatment the ALKO3529 shows the highest increase in lightness unitscompared to VTT-D-79125, ALKO3528, ALKO4097 or ALKO3760 preparations.After 2 hours treatment with 3 mg/g dosage the best stone-washing effectmeasured as lightness units is obtained with ALKO3529 preparation.

EXAMPLE 4

Use of different Trichoderma cellulase preparations in biofinishing ofcotton-containing woven fabric

100 % cotton woven fabric (obtained from Pirkanmaan Uusi Varjaamo Ltd,Finland) was subjected to treatment with VTT-D-79125, ALKO2656,ALKO3529, ALKO3528, ALKO3760, ALKO3798, ALKO4097 cellulase preparations(example 1) in Launder-Ometer. 20 g of undyed prewashed fabric was usedin each experiment. The cellulase treatment conditions were as describedin example 2 except that the liquid ratio (volume of liquid per weightof fabric) was 1:15. 2 and 6 mg of total protein in cellulasepreparations per g of fabric was used in each experiment. The washingtime was 1 hour at pH 5, 50° C. After rinsing with alkaline and water,the treated fabrics were dried in a tumble drier (Cylinda 7703, Sweden).

The following methods were used for evaluation of the effects of thecellulase treatments on cotton fabrics: Weight loss of the treatedfabrics was defined as percentage from weight of the fabric before andafter the test (before weighting the fabrics were conditioning in anatmosphere of 21±2° C. and 50±2% RH). Weight loss was used to describethe amount of fuzz removed from the fabrics surface.

Visual appearance of the enzyme treated fabrics was performed by a panelconsisting of five persons. The fabrics were ranked on a score from 1 to5, where 5 gave a clean surface with no fuzz and pills and the fabrictexture became more apparent. Score of 1 gave many pills and fuzz.

The Martindale Rubbing method (SFS-4328) was used for evaluation ofpilling. Pilling was evaluated on a score from 1 to 5 by a panel after200 cycles of abrasion (1=many pills, 5=no pills). The results are shownin Table III and in FIG. 7.

                  TABLE III    ______________________________________    Weight loss, visual appearance, and pilling of the    fabrics treated in Launder-Ometer with different cellulase    preparations.             dosage  weight loss                               visual  pilling after    preparation             mg/g    %         appearance                                       200 cycles    ______________________________________    --       --      0         1       1    VTT-D-79125             2       0.8       2.3     2.3             6       2.2       2.7     3.6    ALKO3760 2       2.1       3.1     2.3             6       4.2       3.4     3.8    ALKO3798 2       3.1       3.3     3.1             6       4.9       3.4     4.3    ALKO4097 2       0         1.5     1.2             6       1.0       2.5     2.2    ALKO2656 2       2.5       3.8     3.5             6       3.6       3.8     4.4    ALKO3529 2       3.1       3.9     4.3             6       4.8       3.8     4.4    ALKO3528 2       0.7       3.7     3.5             6       2.0       3.4     4.3    ______________________________________

The results show that the best visual appearance and the greatestreduction on pilling tendency is achieved with ALKO3529 cellulasepreparation when comparing the same dosages of different preparations.Also to obtain the same level of pilling removal (e.g. 4.3) ALKO 3529 isneeded in considerably smaller dosages than the other preparations.

EXAMPLE 5

Use of different Trichoderma cellulase preparations in biofinishing ofcotton-containing woven fabric

100% cotton woven fabric (obtained from Pirkanmaan Uusi Varjaamo Ltd,Finland) was subjected to treatment with VTT-D-79125, ALKO3760,ALKO3798, ALKO2656, ALKO3529 and ALKO3528 (example 1) preparations inGavazzi s.r.l. Campiocolor mod. RD/1 (Italy). About 25 g undyedprewashed of fabric and 10 liter of water was used in each experiment atpH 5. The enzyme dosages were 6 and 12 mg of total protein in cellulasepreparations per g of fabric. The washing times was 1 hour at pH 5 50°C.

For evaluation of the effects of the cellulase treatments on cottonfabrics the same methods were used as described in example 4. Thetensile strength was measured according to the SFS Standard 3981.Results are shown in Table VI.

                  TABLE IV    ______________________________________    Weight loss, visual appearance, pilling and tensile    strength of the fabrics treated in Gavazzi Campiocolor with    different cellulase preparations.                                    pilling tensile             dosage  weight   visual                                    after   strength    preparation             mg/g    loss %   appear.                                    200 cycles                                            (weft) N    ______________________________________    --       --      0        1     1       15.5    VTT-D-79125             6       1.9      2.3   3.2     14.0             12      2.6      2.8   3.6     13.4    ALKO3760 6       2.7      3.6   3.2     14.2             12      3.1      3.1   3.8     13.4    ALKO3798 6       2.8      3.0   3.5     13.3             12      3.9      3.8   4.5     12.7    ALKO2656 6       2.3      3.1   3.3     13.8             12      2.5      3.6   3.8     13.8    ALKO3529 6       2.9      4.3   3.7     12.9             12      4.1      4.5   4.4     13.4    ALKO3528 6       1.5      2.9   2.2     13.1             12      2.2      2.9   2.9     13.9    ______________________________________

The best visual appearance and the greatest reduction in pillingtendency was achieved with ALKO3529 cellulase preparation.

EXAMPLE 6

Use of different Trichoderma cellulase preparations in biofinishing ofcotton-containing knit

100% knit (obtained from Apropos Finland, Ltd, Finland) was subjected totreatment with VTT-D-79125, ALKO2656, ALKO3529, ALKO3528, ALKO3760,ALKO3798 and ALKO4097 cellulase preperations (example 1) inLaunder-Ometer. 20 g of undyed knit was used in each experiment and thecellulase treatment conditions were as described in example 2 exceptthat the liquid ratio (volume of liquid per weight of fabric) was 1:15.1, 2 and 4 mg of total protein in cellulase preparations per g of fabricwas used in each experiment. The washing time was 1 hour at pH 5, 50° C.After rinsing with alkaline and water the treated knits were dried intumble drier.

For evaluation of the effects of the cellulase treatments on cottonknits the same methods were used as described in example 4.

                  TABLE V    ______________________________________    Weight loss, visual appearance and pilling of the    cotton knits treated in Launder-Ometer with different cellu-    lase preparations.                       visual pilling after            dosage                  weight loss                             appear-  200  2000    preparation              mg/g    %          ance   cycles    ______________________________________    VTT-D-79125              1       4.4        2.0    1.3  ND              2       5.5        3.3    2.7  1.9              4       6.8        3.5    2.2  ND    average                      2.9    2.1    ALKO3760  1       4.3        2.0    1.7  ND              2       5.4        3.2    2.3  2.5              4       6.8        3.2    1.0  ND    average                      2.8    1.7    ALKO3798  1       4.9        2.3    2.2  ND              2       6.1        2.8    2.2  2.1              4       7.6        3.0    1.8  ND    average                      2.7    2.1    ALKO2656  1       4.4        3.0    1.5  ND              2       6.0        3.8    3.2  2.8              4       6.9        3.7    2.2  ND    average                      3.5    2.3    ALKO3529  1       5.4        3.0    2.0  ND              2       6.7        3.8    3.1  2.9              4       8.4        3.0    2.3  ND    average                      3.3    2.5    ALKO352B  1       4.6        2.7    1.7  ND              2       5.6        3.5    2.1  2.3              4       5.9        3.7    2.0  ND    average                      3.3    1.9    ______________________________________     ND = not done

The results shoe that after 1 hour treatment on the average the best andalmost equal visual appearance was obtained with ALKO2656, ALKO3529 orALKO3528 cellulase treated knits. However on the average the greatestreduction in pilling tendency was obtained with ALK03529 treated knits.

EXAMPLE 7

Use of different Trichoderma cellulase preparations in biofinishing ofcotton-containing knit

100% cotton knit (obtained from Apropos Finland Ltd, Finland) wassubjected to treatment with VTT-D-79125, ALKO2656, ALKO3529, ALKO3528,ALKO3760 and ALKO3798 cellulase preparations (example 1) in Gavazzis.r.l. Campiocolor mod. RD/1 (Italy) About 230 g of knit and 10 liter ofwater was used in each experiment at pH 5. The enzyme dosage was 6 mg oftotal protein in cellulase preparations per g of fabric. The washingtimes was 1 hour at 50° C.

For evaluation of the effects of the cellulase treatments on cottonknits the same methods were used as described in example 4.

Results are shown in Table VI.

                  TABLE VI    ______________________________________    Weight loss, visual appearance and pilling of the    cotton knits treated in Gavazzi Campiocolor with different    cellulase preparations.                            visual                weight loss appear- pilling    preparation %           ance    after 200 cycles    ______________________________________    --          2.7         1       1.1    VTT-D-79125 4.2         3.4     2.3    ALKO3760    4.6         2.6     2.5    ALKO3798    5.2         3.4     3.1    ALKO2656    4.6         3.6     2.8    ALKO3529    5.7         3.8     3.5    ALKO3528    3.6         2.9     2.6    ______________________________________

In Table VI is shown that in treatment of the cotton knit the bestvisual appearance and the greatest reduction in pilling tendency wereachieved with ALKO3529 cellulase preparation.

EXAMPLE 8

Use of different Trichoderma cellulase preparations in biofinishing ofcotton fabric

100% cotton woven fabric (obtained from Pirkanmaan Uusi Varjaamo Ltd,Finland) was subjected to the treatment with ALKO3529, ALKO3760 andALKO3798 cellulase preparations in semi-industrial drum washer, Esteri20 HS-P. About 1.5 kg of fabric was used in each experiment. Waterintake of the machine was about 100 litres. pH was adjusted to 5,treatment time was 45 minutes at 50° C. Enzyme dosage was 6 mg of totalprotein in cellulase preparation per g of fabric.

After enzyme treatment the fabrics were dyed with three differentreactive dyes, Remazol (Bayer), Levafix (Bayer) and Cibacron (Ciba)using normal dying procedures.

The effects of the cellulose treatments on dyed cotton fabrics wereevaluated by the visual appearance of the panel. Results are shown inTable VII.

                  TABLE VII    ______________________________________    The visual appearance of the fabrics treated with    different cellulase preparation in Esteri 20 HS-P washing    machine. Dosage 6 mg total protein/g fabric.    preparation               Remazol      Levafix Cibacron    ______________________________________    --         1.3          1.3     1.3    ALKO3529   4.3          4.1     4.4    ALKO3760   3.8          3.6     3.8    ALKO3798   3.6          3.8     4.0    ______________________________________

The results show that with ALKO3529 the best visual appearance wasobtained on all fabrics.

EXAMPLE 9

Use of different Trichoderma cellulase preparations in biofinishing oflyocell fabric

The ALKO3760, ALKO3798, ALKO4097, ALKO3529, ALKO2656 and ALKO3528(example 1) cellulase preparations were used in fibrillation control andbiofinishing of 100% Tencel® (lyocell). Before enzyme treatment thefabric was prefibrillated in a semi-industrial drum-washer (Esteri 20HS-P) with 2.5 g/l of sodium carbonate at 60° C. The cellulasetreatments were performed in Gavazzi s.r.l. Campiocolor mod. RD/1(Italy). About 70 g of prefibrillated fabric and 10 liter of water wasused in each experiment at pH 5. The enzyme dosages was 6 mg of totalprotein in cellulase preparations per g of fabric. The washing time was2 hours at pH 5 50° C.

The following methods were used for evaluation of the effects of thecellulase treatments on Tencel® fabrics:

* Visual appearance of the enzyme treated fabrics was performed by apanel consisting of five persons. The fabrics were ranked on a scorefrom 1 to 5, where 5 gave a clean surface with no fuzz, fibrils andpills. Score of 1 gave many pills, fibrils and fuzz.

* Fibrillation removal (index): Surface hairs were taken from the fabricand placed in water on a glass slide. This was viewed under the lightmicroscope and Zen areas of the slide were looked at and the resultsaveraged. The smaller the fibrillation index, the more effective enzyme.

Results are shown in Table VIII.

                  TABLE VIII    ______________________________________    The visual appearance and fibrillation index of    the fabrics treated with different cellulase preparations in    Gavazzi Campiocolor.    preparation  visual appearance                             fibrillation index    ______________________________________    --           1           10.2    ALKO3760     3.6         6.1    ALKO3798     3.3         8.1    ALKO4097     3.3         8.0    ALKO2656     3.7         8.2    ALKO3529     4.4         6.8    ALKO3528     3.1         6.4    ______________________________________

Fibrillation index is an average of two analyses. Visual appearance isan average from right and reverse side of the fabrics.

The best visual appearance was obtained with ALKO3529. The fibrillationremoval was most effective with ALKO3760, ALKO3528 and ALKO3529preparations. References referred to in the general description and theexamples are listed in the following:

References:

Bailey, M. and Nevalairien, H. 1981. Induction, isolation and testing ofstable Trichoderma reesei mutants with improved production ofsolubilizing cellulase. Enz. Micr. Tech. 3:153-157.

Benson, S. A. 1984. Bio/Techniques 2:66-68.

Bhat, K. M., McCrae, S. I. and Wood, T. M. 1989. Theendo-1-4-beta-d-glucanase system of penicillium, aspergillus-pinophilumcellulase isolation purification and characterization of five majorendoglucanase components. Carbohydrate Research 190:279-297.

Bhikhabhai, R., Johansson, G. and Pettersson, G. 1984. Isolation ofCellulolytic Enzymes from Trichoderma reesei QM9414. Journal of AppliedBiochemistry 6:335-345.

Buehler, R. 1991. Double-antibody sandwich enzyme-linked immunosorbentassay for quantitation of endoglucanase I of Trichoderma reesei. Appl.Environ. Microbiol. 57:3317-3321.

Drocourt, D., Calmels, T., Reynes, J. P., Baron, M. and Tiraby, G. 1990.Cassettes of the Streptoalloteichus hindustanus ble gene fortransformation of lower and higher eukaryotes to phleomycin resistance.Nuc. Acids Res. 18:4009.

IUPAC Commission on Biotechnology. 1984. Measurement of CellulaseActivities, Ghose, T. K.

Hynes, M., Corrick, C. and King, S. 1983. Isolation of genomic clonescontaining the amdS gene of Aspergillus nidulans and their use in theanalysis of the structural and regulatory mutations. Mo. Cell. Biol.3:1430-1439.

Karhunen, T., Mantyla, A., Nevalainen, H. and Suominen, P. 1993. Highfrequency one-step gene replacement in Trichoderma reesei. I.Endoglucanase I overproduction. Mol. Gen. Genet. 241:515-522.

Kelly, J. and Hynes, M. 1985. Transformation of Aspergillus niger by theamdS gene of Aspergillus nidulans. EMBO J. 4:475-479.

Lowry, O. H., Rosebrough, N. J., Farr, A. L. and Randall, R. J. 1951.Protein measurement with the Folin phenol reagent. J. Biol. Chem.193:265-275.

Mach, R. L., Schindler, M. and Kubicek, C. P. 1994. Transformation ofTrichoderma reesei based on hygromycin B resistance using homologousexpression signals. Curr. Genet. 25:567-570.

Maniatis, T., Fritsch, E. F. and Sambrook, J. 1982. Molecular cloning: Alaboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor,N.Y., USA.

Mattern, J. E., Punt, P. J. and van den Hondel, C. A. M. J. 1988. Avector of Aspergillus transformation conferring phleomycin resistance.Fungal Genet. Newslett. 35:25.

International Patent Publication No. WO 96/34945.

Nevalainen, Genetic improvement of enzyme production in industriallyimportant fungal strains. 1985 VTT, Technical Research Centre ofFinland, publications 26, Espoo Finland.

Penttila et al. 1987. A versatile transformation system for thecellulolytic filamentous fungus Trichoderma reesei. Gene 61:155-164.

Penttila, M., Lehtovaara, P., Nevalainen, H., Bhikhabhai, R. andKnowles, J. 1986. Homology between cellulase genes of Trichodermareesei: complete nucleotide sequence of the endoglucanase I gene. Gene45:253-263.

Saloheimo, M., Lehtovaara, P., Penttila, M., Teeri, T., Stahlberg, J.,Johansson, G., Claeyssens, M., Tomme, P. and Knowles, J. 1988. EGIII, anew endoglucanase from Trichoderma reesei: the characterization of bothgene and enzyme. Gene 63:11-21.

Schulein, M. 1988. Cellulases of Trichoderma reesei. Methods inEnzymology 160:234-242.

Shoemaker, S., Schweikart, V., Ladner, M., Gelfand, D., Kwok, S.,Myambo, K. and Innis, M. 1983. Molecular cloning of exocellobiohydrolasefrom Trichoderma reesei strain L27. Bio/Technology 1:691-696.

Suominen, P., Mantyla, A., Karhunen, T., Hakola, S. and Nevalainen, H.1993. High frequency one-step gene replacement in Trichoderma reesei. IIEffects of deletions of individual cellulase genes. Mol. Gen. Genet.241:523-530.

Teeri, T., Salovuori, I. and Knowles, J. 1983. The molecular cloning ofthe major cellulase gene from Trichoderma reesei. Bio/Technology1:696-699.

Teeri, T., Lehtovaara, P., Kauppinen, S., Salovuori, I. and Knowles, J.1987. Homologous domains in Trichoderma reesei cellulolytic enzymes:gene sequence and expression of cellobiohydrolase II.

van Tilbeurgh, H., Loontiens, F. G., De Bruyne, C. K. and Clayessens, M.1988. Fluorogenic and chromogenic glycosides as substrates and ligandsof carbohydrases. Methods in Enzymology 160:45-59.

Wood, T. M., McCrae, S. I., Wilson, C. A., Bhat, K. M. and Gow, L. A.1988. Aerobic and anaerobic fungal cellulases, with special reference totheir mode of attack on crystalline cellulose. FEMS Symposium No. 43,Biochemistry and Genetics of Cellulose Degradation, pp. 31-52.

    __________________________________________________________________________    SEQUENCE LISTING    (1) GENERAL INFORMATION:    (iii) NUMBER OF SEQUENCES: 2    (2) INFORMATION FOR SEQ ID NO:1:    (i) SEQUENCE CHARACTERISTICS:    (A) LENGTH: 15 base pairs    (B) TYPE: nucleic acid    (C) STRANDEDNESS: single    (D) TOPOLOGY: linear    (ii) MOLECULE TYPE: nucleic acid    (xi) SEQUENCE DESCRIPTION: SEQ ID NO:1:    CCGCGGACTGGCATC15    (2) INFORMATION FOR SEQ ID NO:2:    (i) SEQUENCE CHARACTERISTICS:    (A) LENGTH: 16 base pairs    (B) TYPE: nucleic acid    (C) STRANDEDNESS: single    (D) TOPOLOGY: linear    (ii) MOLECULE TYPE: nucleic acid    (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2:    CCGCGGACTGCGCATC16    __________________________________________________________________________

What we claim:
 1. A method for treating cellulose containing textile material, said method comprising:(i) adding EGII type components to a composition to produce a treatment medium; and (ii) treating said material with said treatment medium.
 2. The method of claim 1, wherein said composition is a natural complete cellulase composition obtained from the fermentation medium of a microbe.
 3. The method of claim 1 or 2, wherein 15-100 weight percent of the total cellulase content of said treatment medium is EGII type components.
 4. The method of claim 3, wherein 20-60 weight percent of the total cellulase content of said treatment medium is EGII type components.
 5. The method of claim 4, wherein 25-40 weight percent of the total cellulase content of said treatment medium is EGII type components.
 6. The method of claim 1, wherein said treatment medium is essentially free of at least one CBH type component.
 7. The method of claim 1, wherein said treatment medium is essentially free of at least one EG type component other than EGII.
 8. The method of claim 1, wherein said treatment medium is essentially free of CBH type components and all EG type components other than EGII.
 9. A method for treating cellulose containing textile materials, said method comprising treating said materials with a treatment medium which comprises a cellulase composition, wherein said cellulase composition is produced by a microbe that has been modified so that said cellulase composition contains an increased weight percent of EGII type components as compared to the weight percent of EGII components in the cellulase composition produced by said microbe prior to said modification.
 10. The method of claim 9, wherein 15-100 weight percent of the total cellulase content of said composition is EGII type components.
 11. The method of claim 10, wherein 20-60 weight percent of the total cellulase content of said composition is EGII type components.
 12. The method of claim 11, wherein 25-40 weight percent of the total cellulase content of said composition is EGII type components.
 13. The method of claim 9, wherein said cellulase composition is essentially free of at least one CBH type component.
 14. The method of claim 9, wherein said cellulase composition is essentially free of at least one EG type component other than EGII.
 15. The method of claim 9, wherein said cellulase composition is essentially free of at least one CBH type component and all EG type components other than EGII.
 16. The method of claim 2 or 9, wherein said microbe is selected from the group consisting of fungi or bacteria.
 17. The method of claim 16, wherein said fungi are selected from the group consisting of Trichoderma, Penicillium, Aspergillus, Humicola and Fusarium genera.
 18. The method of claim 17, wherein said fungi are Trichoderma.
 19. The method of claim 18, wherein said Trichoderma is Trichoderma reesei.
 20. The method of claim 9, wherein said modified microbe is an EGII overproducing Trichoderma reesei strain.
 21. The method of claim 1 or 9, wherein said treatment medium contains surfactants, polymers, buffers, bulking agents, preservatives, stabilizers and/or abrasion agents.
 22. The method of claim 1 or 9, wherein the proportion of cellulosic fiber in the said material is at least 30 percent.
 23. The method of claim 1 or 9, wherein the proportion of cellulosic fiber in said textile material is at least 50 percent.
 24. The method of claim 1 or 9, wherein the cellulosic fiber in said cellulose-containing textile material is selected from a group consisting of cotton, flax, ramie, jute, viscose, modified viscose fibers, lyocell and cupro.
 25. The method of claim 1 or 9, wherein the cellulosic fiber in said cellulose-containing textile material is cotton. 